The major objectives of this research are: 1) to continue our studies to characterize components of specialized cisternae of endoplasmic reticulum which are lined with carbohydrates and which very likely serve as a transport system in neuronal processes, and 2) to continue our studies to relate the localization of macromolecules to the expression of different functions performed by neurons and glia. Macromolecules of nervous tissue membranes will be localized by: 1) the use of a battery of lectins which bind to different sugars and toxins which bind to specific receptors in combination with peroxidase labeling techniques to visualize sites of such binding, and 2) the use of antisera to specific brain macromolecules together with peroxidase conjugated antibodies to visualize tissue sites containing these macromolecules. We will continue to characterize potential candidates for glycoproteins lining the specialized smooth membrane cisternae using lectin affinity chromatography, gel electrophoresis and immunocytochemical procedures. Antisera to those glycoproteins which do line the smooth membrane cisternae will be used in experiments to determine if transport is blocked when specific carbohydrates lining the cisternae are completed with antibody protein. The possible involvement of carbohydrate lined cisternae in transport will be tested further by studying whether or not specific membrane probes will be taken up by neurons and become associated with the cisternae during transport and what factors influence both uptake into the cell and the association of the probes with cisternae. Coated vesicles, another subcellular organelle which may be involved in transport, will be studied using immunocytochemical methods to localize specialized regions of the synaptic terminal formed by the coat protein, and antisera to this protein will be used to see whether or not formation of coated vesicles leading to cellular uptake of macromolecules can be blocked and if such a block leads to inhibition of transport.